• Journal of Microbiology and Biotechnology
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• 제26권3호
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• pp.461-468
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• 2016

In recent years, various naturally occurring defence peptides such as plectasin have attracted considerable research interest because they could serve as alternatives to antibiotics. However, the production of plectasin from natural microorganisms is still not commercially feasible because of its low expression levels and weak stability. A tandemly arrayed plectasin gene (1,002 bp) from Pseudoplectania nigrella was generated using the isoschizomer construction method, and was inserted into the pPICZαA vector and expressed in Pichia pastoris. The selected P. pastoris strain yielded 143 μg/ml recombinant plectasin (Ple) under the control of the methanol-inducible alcohol oxidase 1 (AOX1) promoter. Ple was estimated by SDS-PAGE to be 41 kDa. In vitro studies have shown that Ple efficiently inhibited the growth of several gram-positive bacteria such as Streptococcus suis and Staphylococcus aureus. S. suis is the most sensitive bacterial species to Ple, with a minimum inhibitory concentration (MIC) of 4 μg/ml. Importantly, Ple exhibited resistance to pepsin but it was quite sensitive to trypsin and maintained antimicrobial activity over a wide pH range (pH 2.0 to 10.0). P. pastoris offers an attractive system for the cost-effective production of Ple. The antimicrobial activity of Ple suggested that it could be a potential alternative to antibiotics against S. suis and S. aureus infections.

• Journal of Microbiology and Biotechnology
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• 제13권5호
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• pp.751-758
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• 2003

Mutations in the rdxA gene had been reported to be associated with metronidazole resistance in Helicobacter pylori. In this study, sensitivity to metronidazole, RAPD profiles, and DNA sequences of the rdxA gene of 32 local H. pylori isolates were analyzed. Of these, 13 were found to be resistant, while 19 were sensitive to metronidazole. Among the 32 isolates, 10 were paired isolates from the antrum and body of the stomach of individual patients. Interestingly, the RAPD profiles of isolates from individual patients were distinctly different from each other, whereas paired isolates from the same patient were identical regardless of their sensitivities to metronidazole. DNA sequences of the rdxA gene of all 32 isolates showed 95% to 97% homology when compared with the HP0954 locus of H. pylori 26695 genome. From the 19 metronidazole-sensitive strains, 10 (with $MIC{\le}0.5\;\mu\textrm{g}/ml$ metronidazole) were selected and induced to become metronidazole resistant by sequentially passaging through serial 2-fold increasing concentrations of metronidazole. Nine of the 10 induced paired isolates showed mutations in the rdxA sequences which resulted in truncated protein or changes in the translated amino acid sequences. However, the changes did not occur at any specific site in the DNA or amino acid sequences of the rdxA gene of all the isolates analyzed. The results show that the rdxA gene cannot be a definitive marker for metronidazole resistance in H. pylori isolates of an Asian population, and that other factors may contribute to resistance to metronidazole.

• 한국미생물·생명공학회지
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• 제45권3호
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• pp.209-217
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• 2017

In the enduring investigation of the bioactive microbes, Pseudomonas aeruginosa strain (referred to as CGK-KS-1 (ICTB-315)), isolated from Chumathang hot spring, Ladakh, and India, was identified to possess a major bioactive fraction with antimicrobial and anti-biofilm properties. This bioactive metabolite was purified through bioactivity-guided fractionation. The chemical structure of this major compound was elucidated as phenazine-1-carboxamide (PCN) based on $^1H$ and $^{13}C$ NMR, FT-IR, EI-HR-MS and 2D NMR spectroscopic techniques. In the current study, PCN exhibited antimicrobial activity with MIC values ranging between $1.9-3.9{\mu}g/ml$ against various test human pathogens and Xanthomonas spp. PCN showed the anti-biofilm property with the $IC_{50}$ values ranging from 17.04 to $60.7{\mu}M$ against different test pathogens. The in silico docking studies showed PCN strongly interacted with various proteins of different Xanthomonas spp. with high binding energies. We report herein for the first time the anti-biofilm property and the docking studies of PCN. The extrolite from P. aeruginosa strain CGK-KS-1 showed promising bioactivities and may be considered as a potential candidate for application in various biocontrol strategies.

• 대한수의학회지
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• 제43권4호
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• pp.625-631
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• 2003

The extracts from 40 different traditional medicinal herbs were used to investigate the antimicrobial activities against Escherichia coli. Among them, the extracts from Paeonia suffruticosa (PS), Siegesbeckia orientalis (SO), Schizandra chinensis (SC), Caesalpinia sappan (CS) and Rhus javanica (RJ) exhibited high antimicrobial activities against E. coli, Minimum inhibitory concentrations (MIC) of the RJ extract against E. coli were 0.8 mg/ml. After heating treatment of these extracts, the antimicrobial activities against E. coli were significantly reduced in case of the CS extract. After alkaline or acid treatment of these extracts, the antimicrobial activities against E. coli were significantly increased in the PS extract but reduced in both SO and RJ extract. Since extracts from RJ and CS exhibited the highest antimicrobial activities, bacterial growth-inhibiting activities against E. coli by these two extracts were further examined. Optical density at 620 nm after 24 hours incubation of E. coli in the presence of 100, 300 or 500 ppm of RJ extract ranged from 0.1 to 0.2 compared to 0.35~0.65 in the absence of RJ extract, indicating that growth of E. coli was significantly inhibited within 24 hours by the addition of at least 100 ppm of RJ extract. Optical density at 620 nm after 24 hours incubation of E. coli in the presence of 300 or 500 ppm of CS extract ranged from 0.01 to 0.25 compared to 0.5~0.55 in the absence of CS extract, indicating that growth of E. coli was also significantly inhibited within 24 hours by the addition of at least 300 ppm of CS extract. In conclusion, these findings suggest that extracts from RJ and CS may play important roles for antimicrobial activities against E. coli causing various animal diseases.

• Bulletin of the Korean Chemical Society
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• 제32권8호
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• pp.2695-2699
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• 2011

${\beta}$ Ketoacyl acyl carrier protein synthase III (KAS III) initiates fatty acid synthesis in bacteria and is a key target enzyme to overcome the antibiotic resistance problem. In our previous study, we found flavonoid inhibitors of Enterococcus faecalis KAS III and proposed three potent antimicrobial flavonoids against Enterococcus faecalis and Vancomycin-resistant Enterococcus faecalis with MIC values in the range of 128-512 ${\mu}g/mL$ as well as high binding affinities on the order from $10^6$ to $10^7\;M^{-1}$. Using these series of flavonoids, we conducted biological assays as well as docking study to find potent flavonoids inhibitors of Staphylococcus aureus KAS III with specificities against Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. Here, we propose that naringenin (5,7,4'-trihydroxyflavanone) and eriodictyol (5,7,3',4'-tetrahydroxyflavanone) are potent antimicrobial inhibitors of Staphylococcus aureus KAS III with binding affinity of $3.35{\times}10^5$ and $2.01{\times}10^5\;M^{-1}$, respectively. Since Arg38 in efKAS III is replaced with Met36 in saKAS III, this key difference caused one hydrogen bond missing in saKAS III compared with efKAS III, resulting in slight discrepancy in their binding interactions as well as decrease in binding affinities. 4'-OH and 7-OH of these flavonoids participated in hydrogen bonding interactions with backbone carbonyl of Phe298 and Ser152, respectively. In particular, these flavonoids display potent antimicrobial activities against various MRSA strains in the range of 64 to 128 ${\mu}M$ with good binding affinities.

• KSBB Journal
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• 제10권2호
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• pp.131-136
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• 1995

본 실험에서는 인간 전과립 세포를 회분배양 조건 에서 10%의 혈청을 포함하는 배지로 배양하였다. 이 와같은 조건에서 세포의 비성장율은 0.374C1/day) 이고 세포내 존재하는 살균활성을 가진 목적 단백질 의 생산성은 0.435(mg/10' viable cells)으로 나타났 다. 세포내 존재하는 목척 단백질의 순수 분리를 위 해 sephadex G-75, sephadex G-100, Bio→Rex70 을 이용해 분리하였다. 정제된 목적 단백질은 감수성 균주, E. coli IFO 13168과 St. aureus IFO 3060 을 표준 균주로 살균 측정을 하였을 때, 기존의 항생 물질은 균주에 대해 40~701땅1m!에서 활성을 나타 낸 반면, 정제된 목적 단백질은 G(+)균(St. aureus IFO 3060)에서는 $0.5\mu\textrm{g}$/ml이고, G(-)균(E. coli IFO 13168)에서는 $0.4\mu\textrm{g}$/ml에서 살균 활성을 보였다. 따라서 AMF가 기존 항생물질보다 살균 활성이 강력한 단백질임을 알 수 있다. 해당 분자량을 측정 하기 위해 SDS-PAGE로 측정한 결과 약 15,000 Dalton의 분자량을 갖고 있음이 확인됐다.

• 대한화장품학회지
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• 제25권4호
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• pp.111-121
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• 1999

Acne vulgaris, the most common skin disease. can be formed as only a few comedons or severe inflammatory lesions. The pathogenesis of acne involves various factors; excessive androgen, excessive sebum production, abnormal alteration of follicular epithelium, proliferation of Propionibacterium acnes, and inflammation. We investigated acne therapy using oriental herbs described in the Korean traditional medical book(Dong-ui-bo-gam). Oriental herbs(Angelica daurica, Arctium lappa, Coptidis rhizoma, and Glycyrrhiza glabra) were chosen based on their respective property of sebum control, anti-inflammatory activity, and anti-bacterial activity. We examined the effect of acne treatment, in terms of chemotactic inhibition, lipogenesis inhibition, and anti-bacterial activity for P. acnes. 1. Neutrophil chemotaxis assay; P. acnes secrete chemotactic factors and other pro-inflammatory extracellular products. Neutrophil chemotactic activity of P. acnes was measured by 48-well chemotaxis method. Angelica daurica clearly suppressed chemotactic activity of P. acnes. 2. Using sebaceous gland of hamster ear lipogenesis assay; Sebaceous lipogenesis was measured using ear biopsies by incubation or $C^{14}$-acetate in culture media. The $C^{14}$-labeled lipids were extracted and determined by liquid scintilation counting. Coptidis rhizoma markedly inhibited sebum production. 3. Anti-bacterial assay for P. acnes(MIC test); Glycyrrhiza glabra showed anti-bacterial activity. P. acnes did not develop resistance against Glycyrrhiza glabra. Retinoids are effectively to inhibit sebum production and regulate follicular keratinization process, with little anti-inflammatory activity. Angelica daurica suppressed neutrophil chemotaxis, Coptidis rhizoma inhibited sebum production, and Glycyrrhiza glabra showed anti-bacterial activity against P. acnes. A combined formulation of Angelica daurica, Coptidis rhizoma. and Glycyrrhiza glabra is expected to provide effective acne treatment.

• 생약학회지
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• 제45권2호
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• pp.168-173
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• 2014

The aim of this study was to evaluate the anti-helicobacter activity of the ethanol extract of Sohamhyoongtang (Coptidis Rhizoma, Pinelliae Tuber and Trichosanthis Semen) and Coptidis Rhizoma total alkaloids, which is one of the components of Sohamhyoongtang. Crude ethanol extract of Sohamhyoongtang (ESHHT) and Coptidis Rhizoma total alkaloids (CRTA) were used for this experiment. Five different types of H. pylori (including H. pylori 26695) were used as test strain. To determine anti-helicobacter activity, minimum inhibitory concentration (MIC) was determined by agar dilution method. The effect of ESHHT and CRTA on the gene expression of H. pylori was investigated by quantitative realtime-PCR (qRT-PCR). MICs of ESHHT against five H. pylori strains were $250{\sim}500{\mu}g/ml$ and MICs of CRTA against five H. pylori strains were $50{\sim}200{\mu}g/ml$. Four representative virulence genes of H. pylori, cagA, ureA, ureB and ureI were tested as target genes for qRT-PCR. According to the qRT-PCR results, both ESHHT and CRTA markedly repressed the expression of cagA gene of H. pylori 26695 (6.91 and 20 folds respecively). These results showed that the ESHHT and CRTA demonstrated antihelicobacter properties, suggesting their potential use in gastritis or duodenal ulcer.

• Asian-Australasian Journal of Animal Sciences
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• 제31권10호
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• pp.1627-1634
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• 2018

Objective: The study aimed to evaluate the effect of replacing wheat bran for cactus cladodes plus urea (0%, 25%, 50%, 75%, and 100%) on the intake of nutrients, nitrogen balance, microbial protein synthesis, and rumen fermentation for steers. Methods: Five crossbred steers (1/2 Holstein-Zebu), with rumen cannula and an average body weight of $180{\pm}5.3kg$, were assigned to a $5{\times}5$ Latin square design. Dietary treatments consisted of the replacement of the total of wheat bran in basal diet by cactus cladodes using the following proportions: 0% for basal diet, 25%, 50%, 75%, and 100% cactus cladodes replacing wheat bran. Urea was added to the diets to adjust the crude protein (CP) content to 130 g/kg dry matter. Results: Maximum dry matter intake (5.73 kg/d) and maximum nitrogen balance (103 g/d) were estimated for 54.6% and 70.8% replacement levels of wheat bran. The maximum microbial protein production (44.6 g/d) was obtained at a replacement level of 49.7%, and a medium value (125 g CP mic/kg total digestible nutrients) of microbial protein efficiency was observed. The rumen pH increased linearly according to cactus cladodes inclusion, while the ammonia nitrogen medium value was 24.5 mg/dL. Conclusion: The replacement of 55% wheat bran for cactus cladodes plus urea in the diet of crossbred steers is recommended.

• 한국수산과학회지
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• 제43권6호
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• pp.642-647
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• 2010

This study examined the antimicrobial activity of Myagropsis yendoi (MY). The antimicrobial activity of ethanol and aqueous MY extracts were measured using a paper disc assay. The MY ethanol extract had the best antimicrobial activity. It inhibited the growth of Candida tropicalis and Gram-positive bacteria such as Bacillus subtilis, Listeria innocua, L. monocytogenes, and Staphylococcus aureus at 4 mg/mL. The minimum inhibitory concentration (MIC) of the MY ethanol extract ranged from 0.1 to 0.5%. The MY ethanol extract inhibited the growth of B. subtilis and L. innocuaat concentrations of 0.15% and 0.1%, respectively. The antimicrobial activity of the MY ethanol extract was not affected by heating at $121^{\circ}C$ for 15 min or pH 2.10. Therefore, the main substances responsible for the antimicrobial activity of the MY ethanol extract are believed to be stable with changing heat and pH.